Nanosensor methods and apparatuses for determination of analytes

Abstract

The present invention generally relates, in some aspects, to articles and methods relating to nanosensors for determination of molecules and other features, e.g., via surface plasmonic resonance, electric resonance, magnetic resonance, color changes, or the like. These articles and methods may be used, for example, for sample detection. The articles described in some aspects of the invention include a microwell array and a nanosensor array. In some embodiments, The nanosensor arrays may utilize nanoparticles positioned on nanostructures that are able to interact with a sample suspected of containing an analyte, such as a single cell. The interaction between nanoparticles and a sample can be detected by a change in applied energy, such as altered electromagnetic radiation caused by surface plasmonic resonance of incident visible light, and/or other types of resonance. Electromagnetic radiation may be applied to a microwell array and nanosensor, and the applied electromagnetic radiation may be altered as a nanosensor interacts with a sample suspected of containing an analyte.

What is claimed is: 
     
       1. A method, comprising:
 allowing a lysing reagent to enter a well of a microwell array through a semipermeable membrane sealing the well, wherein the semipermeable membrane is configured to prevent a lysate from leaving the well; 
 lysing a cell within the well to release an analyte suspected of being able to bind a reaction entity; 
 applying electromagnetic radiation to a nanoparticle at least partially coated with the reaction entity, wherein: (i) the nanoparticle is configured to interact with the electromagnetic radiation via electric resonance and/or magnetic resonance to alter the electromagnetic radiation; and (ii) the well comprises a nanostructure and the nanoparticle is positioned distally on an end of the nanostructure; and 
 determining the altered electromagnetic radiation. 
 
     
     
       2. A method, comprising:
 acquiring a first optical color image of an array of nanostructures on a substrate, wherein the nanostructures have a cross-sectional dimension, orthogonal to the direction that the first optical image is acquired, of less than 700 nm, wherein: (i) the nanostructures are at least partially coated with a reaction entity; (ii) the substrate comprises a microwell array comprising a plurality of wells, each well comprising a nanostructure of the array of nanostructures; and (iii) the nanostructures are attached to the substrate; 
 allowing a lysing reagent to enter a well of the plurality of wells through a semipermeable membrane sealing the well, wherein the semipermeable membrane is configured to prevent a lysate from leaving the well; 
 lysing a cell within the well to release an analyte suspected of being able to bind the reaction entity; 
 causing an interaction between the reaction entity and the analyte; 
 acquiring a second optical color image of the array of nanostructures; and 
 determining a change in color between the first optical image and the second optical image, wherein the change in color is caused by the interaction between the reaction entity and the analyte. 
 
     
     
       3. The method of  claim 1 , wherein the semipermeable membrane comprises polycarbonate. 
     
     
       4. The method of  claim 1 , wherein the semipermeable membrane has a pore size of 10 nm. 
     
     
       5. The method of  claim 1 , wherein the electromagnetic radiation comprises visible light. 
     
     
       6. The method of  claim 1 , further comprising determining the reaction entity by the determined altered electromagnetic radiation. 
     
     
       7. The method of  claim 2 , wherein the array of nanostructures has an average spacing between nanostructures of less than about 3 micrometers. 
     
     
       8. The method of  claim 2 , wherein the array of nanostructures has an average spacing between nanostructures of less than about 500 nm. 
     
     
       9. The method of  claim 2 , wherein the reaction entity comprises an antibody. 
     
     
       10. The method of  claim 2 , wherein the reaction entity comprises an aptamer. 
     
     
       11. The method of  claim 2 , wherein the reaction entity comprises a protein. 
     
     
       12. The method of  claim 2 , wherein the reaction entity comprises an oligonucleotide. 
     
     
       13. The method of  claim 2 , wherein the reaction entity comprises an enzymatic reaction product induced by a chromogenic substrate labeled on the analyte. 
     
     
       14. The method of  claim 13 , wherein the chromogenic substrate is 3,3′,5,5′-tetramethylbenzidine. 
     
     
       15. The method of  claim 13 , wherein the chromogenic substrate is 3,3′-diaminobenzidine. 
     
     
       16. The method of  claim 13 , wherein the chromogenic substrate is 2,2′-azino-di-(3-ethylbenzothiazoline-6-sulfonic acid). 
     
     
       17. The method of  claim 2 , wherein the nanostructure has a length of less than about 5 micrometers. 
     
     
       18. The method of any one of  claims 2 , wherein the nanostructure has a length of at least about 0.01 micrometers. 
     
     
       19. The method of  claim 2 , wherein the nanostructure comprises a semiconductor. 
     
     
       20. The method of  claim 2 , wherein the nanostructure comprises silicon. 
     
     
       21. The method of  claim 2 , wherein the nanostructure consists essentially of silicon.